人结直肠癌干细胞微球体培养及鉴定.PDF

( ) 　2009;40 (5):918 922　　　　　　　　　　　　　　　　　　　　　　　　 J Sichuan Univ (Med Sci Edi) 1 2 1 1 1 1 1 1■ 周继陶 , 龚日祥 , 陈向征 , 周海俊 , 朱亚杰 , 刘素蕊 , 黄　娟 , 毕　锋 1. ( 610041);2. 　　【】　　, Colo205 。　Lovo、Colo205 SW480 (SFM)。 、、Transwell Colo205 CD133、CD44 , 。CD44 。 RT-PCR Colo205CD44、Musashi-1 Oct4 mR A 。　3 SFM , SFM 、 Accutase + 。Colo205 CD44 , Colo205 。Colo205 , , Musashi-1 Oct4 mR A。　SFM , 。 【】　　　　　 【】　735.34 Cultivation and Characterization of Tumor Sphere Generated from Human Colorectal Cancer Cell Line 　Z HOU J i- tao, GON G R i-x iang, CHEN X iang-z heng, Z HOU Hai-j un, Z HU Ya-j ie, LIU S u-rui, H UA N G J uan, I ■ Feng .　Dep artment of Abdomina l Cancer, Cancer Center, West China Hosp ital, S ichuan University , Chengdu 610041, China 【Ab tract】　Objective　To study on the cultivation method for tumor spheres from colorectal cancer cell lines and identify whether resulting Colo205 spheroid cells display cancer stem cell characteristics.Method 　 Lovo, Colo205 and SW480 cells were seeded in serum free medium (SFM)with EGF and bFGF.Flow cytometry analysis, cell invasion assay and xenograft experiment were applied to examine the cell surface marker expression pattern, cell invasive ability and in vivo tumorigenicity of both Colo205 spheres and parental cells.CD44 expression of tumor spheroid cells was also analyzed after cultured with serum supplemented medium by flow cytometry.CD44, Musashi-1 and Oct4 mR A were detected in these two cells by RT-PCR.Re ult 　 Tumor spheres could be generated from three colorectal cancer cell lines in SFM.The formation and proliferation of tumor spheres were benefited from fresh SFM, cell dissociation reagent Accutase a